Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S139) 抗体 [EP656Y] (ab40795)

製品の概要

  • 製品名Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S139) antibody [EP656Y]
    PAK1+PAK2+PAK3 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [EP656Y] to PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S139)
  • 特異性ab40795 recognises p21-activated kinase 1 (PAK1).
  • アプリケーション適用あり: WB, IHC-P, IP, Flow Cyt, ICC/IFmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human PAK1. A synthesized phosphopeptide derived from human PAK1 (SwissProt=Q13153) + PAK2 (SwissProt=Q13177) + PAK3 (SwissProt=O75914) around the phosphorylation sites of serine 144 + 141 + 139.

  • ポジティブ・コントロール
    • WB: HeLa, RAW264.7 and C6 cell lysates. IHC: Human liver carcinoma, mouse cerebral cortex, rat cerebral cortex. ICC/IF, IP: HeLa cell lysate. Flow Cyt: NIH/3T3 cell lysate.
  • 特記事項

    This product is a recombinant rabbit monoclonal antibody.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Alternative versions available:
    Anti-PAK1 + PAK2 + PAK3 (phospho S144, S141, S139) antibody (Alexa Fluor® 647) [EP656Y] (ab203640)
    Anti-PAK1+PAK2+PAK3 (phospho S139 + S141 + S144) antibody (Alexa Fluor® 488) [EP656Y] (ab203958)

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab40795 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
WB 1/10000 - 1/50000. Detects a band of approximately 66 kDa (predicted molecular weight: 65 kDa).
IHC-P 1/100 - 1/500.
IP 1/40.
Flow Cyt 1/120.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF 1/250 - 1/500.

ターゲット情報

Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S139) antibody [EP656Y] 画像

  • All lanes : Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S139) antibody [EP656Y] (ab40795) at 1/2000 dilution (unpurified)

    Lane 1 : HeLa cell lysate with None
    Lane 2 : HeLa cell lysate with PAK2 (pS141)
    Lane 3 : HeLa cell lysate with PAK2 non-phospho
    Lane 4 : HeLa cell lysate with PAK3 (pS139)
    Lane 5 : HeLa cell lysate with PAK3 non-phospho


    Predicted band size : 65 kDa
  • ab40795 staining PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S139 in rat cerebral cortex tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

    Negative control 1: PBS in place of primary

  • ab40795 staining PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S139) in HeLa (human cervix adenocarcinoma) cells, treated and untreated with Lambda Protein Phosphtase 31℃ for 5h by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody. ab7291 and ab150120 were used as counterstains for primary antibody ab75748 and secondary antibody ab150077 respectively and DAPI was used as a nuclear counterstain.

    Negative control 1: Rabbit primary antibody and anti-mouse secondary antibody (ab150120)
    Negative control 2: Mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077)

     

  • ab40795 staining PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S139) in the human cell line NIH/3T3 (mouse embryo) by flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/120. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/500 was used as the secondary antibody.

    Isoytype control: Rabbit monoclonal IgG (Black)

    Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

     

  • ab40795 immunoprecipitating PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S139). 10µg of cell lysate was incubated with primary antibody at a dilution of 1/40 and VeriBlot for IP secondary antibody (HRP) (ab131366) at a dilution of 1/10000.

    Lane 1: HeLa (human cervix adenocarcinoma) whole cell lysate (10ug)
    Lane 2: HeLa (human cervix adenocarcinoma) whole cell lysate
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab40795 in HeLa (human cervix adenocarcinoma) whole cell lysate

     

  • All lanes : Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S139) antibody [EP656Y] (ab40795) at 1/50000 dilution

    Lane 1 : C6 (rat glioma) whole cell lysate . The membrane is then treated with phosphatase.
    Lane 2 : Untreated C6 (rat glioma) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

    Predicted band size : 65 kDa
  • ab40795 staining PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S139 in mouse cerebral cortex tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

    Negative control 1: PBS in place of primary antibody.

  • Overlay histogram showing HeLa cells stained with unpurified ab40795 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab40795, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

  • All lanes : Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S139) antibody [EP656Y] (ab40795) at 1/10000 dilution

    Lane 1 : Untreated HeLa (human cervix adenocarcinoma) whole cell lysate
    Lane 2 : HeLa (human cervix adenocarcinoma) whole cell lysate. The membrane is then treated with phosphatase.

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution

    Predicted band size : 65 kDa
  • ab40795 staining PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S139 in human liver carcinoma tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

    Negative control 1: PBS in place of primary antibody.

     

  • All lanes : Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S139) antibody [EP656Y] (ab40795) at 1/10000 dilution

    Lane 1 : RAW264.7 (mouse abelson murine leukemia virus-induced tumor) whole cell lysate. The membrane is then treated with phosphatase.
    Lane 2 : Untreated RAW264.7 (mouse abelson murine leukemia virus-induced tumor) whole cell lysate.

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

    Predicted band size : 65 kDa

Anti-PAK1 (phospho S144) + PAK2 (phospho S141) + PAK3 (phospho S139) antibody [EP656Y] (ab40795) 使用論文

This product has been referenced in:
  • Freeman MC  et al. Coronaviruses induce entry-independent, continuous macropinocytosis. MBio 5:e01340-14 (2014). Mouse . Read more (PubMed: 25096879) »
  • El-Baba C  et al. Thymoquinone-induced conformational changes of PAK1 interrupt prosurvival MEK-ERK signaling in colorectal cancer. Mol Cancer 13:201 (2014). WB ; Human . Read more (PubMed: 25174975) »

See all 11 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HCT116)
Loading amount 40 µg
Specification HCT116
Gel Running Conditions Non-reduced Denaturing (15%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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投稿 Nov 30 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (normal tissue array (including breast/colon/spleen)
Specification normal tissue array (including breast/colon/spleen
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate
Permeabilization No
Blocking step Serum as blocking agent for 10 minute(s) · Concentration: 10%
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投稿 May 19 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Breast)
Loading amount 20 µg
Specification Breast
Gel Running Conditions Non-reduced Non-Denaturing (Native) (4-12)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10%
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Verified customer

投稿 May 18 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (cultured hippocampal astrocytes)
Specification cultured hippocampal astrocytes
Fixative Paraformaldehyde
Blocking step BSA as blocking agent for 20 minute(s) · Concentration: 1%
Username

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Verified customer

投稿 Mar 08 2007

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"